Life in extraterrestrial environments Quarterly status report, 1 Sep. - 30 Nov. 1967
Importance of Bacillus cereus and Staphylococcus aureus survival and growth to extraterrestrial planetary contamination
Hagen, C. A.↗
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Life in extraterrestrial environments Quarterly status report, 29 Feb. - 15 May 1968
Incubation temperature and salt concentration effects on Staphylococcus aureus growth in simulated Mars environment
Source record↗
Bacterial Identification Using Light Scattering Measurements: a Preliminary Report
The light scattering properties of single bacterial cells were examined as a possible means of identification. Three species were studied with streptococcus faecalis exhibiting a unique pattern; the light-scattering traces for staphylococcus aureus and escherichia coli were quite similar although differences existed. Based on preliminary investigations, the light scattering approach appeared promising with additional research needed to include a wide variety of bacterial species, computer capability to handle and analyze data, and expansion of light scattering theory to include bacterial cells.
Wilkins, J. R.↗
Effects of mutation and some environmental factors on the physiology and pathogenicity of selected bacteria
Studies with mutants of Staphylococcus aureus lacking some virulence factors suggest that the presence of deoxyribonuclease correlates with mouse pathogenicity of S. aureus, while the ability to ferment mannitol or the possession of coagulases are not required for virulence. Autotrophy investigations on mycobacteria demonstrate a complete correlation between the ability to grow with hydrogen and the species of scotochromogenic mycobacterium tested. All tested strains of M. gordonae, a saprophyte, could grow autotrophically while none of the tested strains of M. scrofulaceum, a clinically important species, possessed this ability. A series of heat tolerant mutants of Pseudomonas fluorescences were obtained which can grow at temperatures up to 54 C, in contrast to a maximum growth temperature of 37 C for the wild type.
Decicco, B. T.↗
Recovery of medically important microorganisms from Apollo astronauts
Microbiological samples were obtained from the crewmembers of the Apollo 13, 14, 15, 16, and 17 spaceflights. These specimens were analyzed for the presence of medically important microorganisms with Staphylococcus aureus, Pseudomonas aeruginosa, Tricophyton mentagrophytes, Tricophyton rubrum, and Candida albicans being discussed in detail. Preflight isolation of crewmembers was found to coincide with a complete absence of inflight disease events and is recommended for future spaceflights. No autoinfection response (microbial shock) occurred after any of the reported spaceflights.
Taylor, G. R.↗
Microbial exchange experiment AR-002
Operational aspects associated with the experiment and the activities of medically important microorganisms recovered from the Apollo crewmen are evaluated. A variety of potential pathogens was recovered from each of the prime and backup crew members before and after flight. However, no disease events were reported. Candida albicans and Staphylococcus aureus were shown to be transferred from one crewmember to another during the flight. No other medically significant changes in the microbial population were observed.
Taylor, G. R.↗
Ecology and thermal inactivation of microbes in and on interplanetary space vehicle components
The thermal resistance of vegetative cells in the percent RH range of 0.19 to 100 at 60 C was investigated. Staphylococcus aureus was used in the experiment because of the extension of the moisture range that can be examined.
Reyes, A. L.↗
Storage Stability and Improvement of Intermediate Moisture Foods, Phase 3
Methods were determined for the improvement of shelf-life stability of intermediate moisture foods (IMF). Microbial challenge studies showed that protection against molds and Staphylococcus aureus could be achieved by a combination of antimicrobial agents, humectants and food acids. Potassium sorbate and propylene glycol gave the best results. It was also confirmed that the maximum in heat resistance shown by vegetative pathogens at intermediate water activities also occurred in a solid food. Glycols and sorbitol both achieve browning inhibition because of their action as a medium for reaction and effect on viscosity of the adsorbed phase. Chemical availability results showed rapid lysine loss before visual discoloration occurred. This is being confirmed with a biological test using Tetrahymena pyriformis W. Accelerated temperature tests show that effectiveness of food antioxidants against rancidity development can be predicted; however, the protection factor changes with temperature. BHA was found to be the best antioxidant for iron catalyzed oxidation.
Labuza, T. P.↗
Methods for microbiological and immunological studies of space flight crews
Systematic laboratory procedures compiled as an outgrowth of a joint U.S./U.S.S.R. microbiological-immunological experiment performed during the Apollo-Soyuz Test Project space flight are presented. Included are mutually compatible methods for the identification of aerobic and microaerophilic bacteria, yeast and yeastlike microorganisms, and filamentous fungi; methods for the bacteriophage typing of Staphylococcus aureus; and methods for determining the sensitivity of S. aureus to antibiotics. Immunological methods using blood and immunological and biochemical methods using salivary parotid fluid are also described. Formulas for media and laboratory reagents used are listed.
Taylor, G. R.↗
Response of selected microorganisms to experimental planetary environments
Anaerobic and aerobic sporeformers and non-sporeformers were cultivated anaerobically in nutrient media under various pressures (up to 1800 psi) of pure H2, CH4, NH3, and H2S. Viability assays were performed periodically to determine growth, survival, or spore survival. Hydrogen up to 1800 psi demonstrated little or no suppression of growth with the possible exception of Bacillus coagulans at 1800 psi. The obligate anaerobes grew very well. Under CH4 the obligate anaerobes again exhibited the most prolific growth, whereas the facultative anaerobes grew well except under higher pressures. Ammonia at low pressure was extremely toxic to all test organisms. At 100 psi all populations were killed within 24 hours except Staphylococcus aureus which survived for 72 hours and the Bacillus spp. which produced a surviving population of approximately 10,000 spores/ml. All populations in H2S were killed within 24 to 48 hours except Proteus mirabilis which decreased to 100 cells/ml and the Bacillus spp. Spore survival studies of two months duration demonstrated that B. coagulans and B. pumilus survived under all experimental conditions. Clostridium novyi type B and C. sporogenes were killed rapidly in NH3 and H2S and demonstrated no sporulation.
Foster, T. L.↗
Preflight and postflight microbiological results from 25 space shuttle crews
Clinical-microbiological investigations are an important aspect of the crew health stabilization program. To ensure that space crews have neither active nor latent infections, clinical specimens, including throat and nasal swabs and urine samples, are collected at 10 days (L-10) and 2days (L-2) before launch, and immediately after landing (L+0). All samples are examined for the presence of bacteria and fungi. In addition, fecal samples are collected at L-10 and examined for bacteria, fungi and parasites. This paper describes clinical-microbiological findings from 144 astronauts participating in 25 Space Shuttle missions spanning Space Transportation System (STS)-26 to STS-50. The spectrum of microbiological findings from the specimens included 25 bacterial and 11 fungal species. Among the bacteria isolated most frequently were Staphylococcus aureus, Enterobacter aerogenes, Enterococcus faecalis, Escherichia coli, Proteus mirabilis and Streptococcus agalactiae. Candida albicans was the most frequently isolated fungal pathogen.
Pierson, Duane L.↗
Survival of Potentially Pathogenic Human-Associated Bacteria in the Rhizosphere of Hydroponically Grown Wheat
Plants may serve as reservoirs for human-associated bacteria (H-AB) in long-term space missions containing bioregenerative life support systems. The current study examined the abilities of five human-associated potential pathogens, Pseudomonas aeruginosa, Pseudomonas cepacia, Staphylococcus aureus, Streptococcus pyogenes, and Escherichia coli, to colonize and grow in the rhizosphere of hydroponically grown wheat, a candidate crop for life support. All of these bacteria have been recovered from past NASA missions and present potential problems for future missions. The abilities of these organisms to adhere to the roots of axenic five-day-old wheat (Triticum aestivum L. cv. Yecora rojo) were evaluated by enumeration of the attached organisms after a one hour incubation of roots in a suspension (approximately 10(exp 8 cu/ml)) of the H-AB. Results showed that a greater percentage of P. aeruginosa cells adhered to the wheat roots than the other four H-AB. Similarly incubated seedlings were also grown under attempted axenic conditions for seven days to examine the potential of each organism to proliferate in the rhizosphere (root colonization capacity). P. cepacia and P. aeruginosa showed considerable growth. E. coli and S. aureus showed no significant growth, and S. pyogenes died off in the wheat rhizosphere. Studies examining the effects of competition on the survival of these microorganisms indicated that P. aeruginosa was the only organism that survived in the rhizosphere of hydroponically grown wheat in the presence of different levels of microbial competition.
Morales, Anabelle↗
Macrophage cell lines derived from major histocompatibility complex II-negative mice
Two bone-marrow-derived macrophage cell lines, C2D and C2Dt, were isolated from major histocompatibility class II negative knock-out mice. The C2D cell line was stabilized by continuous culture in colony-stimulating factor-1 and the C2Dt cell line was transformed with SV40 virus large T antigen. These cells exhibited phenotypic properties of macrophages including morphology and expression of Mac 1 and Mac 2 cell surface molecules. These cells also had comparable growth to the bone-marrow-derived macrophage cell line B6MP102. These new cell lines were not spontaneously cytotoxic and were only capable of modest killing of F5b tumor cells when stimulated with LPS and interferon-gamma, but not when stimulated with LPS alone or with staphylococcal exotoxin. C2D and C2Dt cells phagocytosed labeled Staphylococcus aureus similarly to B6MP102 cells but less well than C2D peritoneal macrophages. These cell lines secreted interleukin-6, but not tumor necrosis factor or nitric oxide in response to LPS or staphlococcal enterotoxins A or B C2D(t) cells were tumorigenic in C2D and C57BL/6J mice but C2D cells were not. These data suggest that macrophage cell lines can be established from bone marrow cells of major histocompatibility complex II-negative mice.
NASA Discipline Musculoskeletal↗
Mouse infection models for space flight immunology
Several immunological processes can be affected by space flight. However, there is little evidence to suggest that flight-induced immunological deficits lead to illness. Therefore, one of our goals has been to define models to examine host resistance during space flight. Our working hypothesis is that space flight crews will come from a heterogeneous population; the immune response gene make-up will be quite varied. It is unknown how much the immune response gene variation contributes to the potential threat from infectious organisms, allergic responses or other long term health problems (e.g. cancer). This article details recent efforts of the Kansas State University gravitational immunology group to assess how population heterogeneity impacts host health, either in laboratory experimental situations and/or using the skeletal unloading model of space-flight stress. This paper details our use of several mouse strains with several different genotypes. In particular, mice with varying MHCII allotypes and mice on the C57BL background with different genetic defects have been particularly useful tools with which to study infections by Staphylococcus aureus, Salmonella typhimurium, Pasteurella pneumotropica and Ehrlichia chaffeensis. We propose that some of these experimental challenge models will be useful to assess the effects of space flight on host resistance to infection.
Review↗
Characterization of Volume F Trash from the Three FY11 STS Missions: Trash Weights and Categorization and Microbial Characterization
The project reported here provides microbial characterization support to the Waste Management Systems (WMS) element of NASA's Life Support and Habitation Systems (LSHS) program. Conventional microbiological methods were used to detect and enumerate microorganisms in STS Volume F Compartment trash for three shuttle missions: STS 133, 134, and 135. This trash was usually made available within 2 days of landing at KSC. The Volume F bag was weighed, opened and the contents were cataloged and placed into categories: personal hygiene items - inclUding EVA maximum absorbent garments (MAGs) and Elbow packs (daily toilet wipes, etc), drink containers, food waste (and containers), office waste (paper), and packaging materials - plastic film and duct tape. The average wet trash generation rate for the three STS missions was 0.362 % 0.157 kgwet crew 1 d-1 . This was considerably lower and more variable than the average rate for 4 STS missions reported for FY10. Trash subtotals by category: personal hygiene wastes, 56%; drink items, 11 %; food wastes, 18%; office waste, 3%; and plastic film, 12%. These wastes have an abundance of easily biodegraded compounds that can support the growth of microorganisms. Microbial characterization of trash showed that large numbers of bacteria and fungi have taken advantage of this readily available nutrient source to proliferate. Exterior and interior surfaces of plastic film bags containing trash were sampled and counts of cultivatable microbes were generally low and mostly occurred on trash bundles within the exterior trash bags. Personal hygiene wastes, drink containers, and food wastes and packaging all contained high levels of, mostly, aerobic heterotrophic bacteria and lower levels of yeasts and molds. Isolates from plate count media were obtained and identified .and were mostly aerobic heterotrophs with some facultative anaerobes. These are usually considered common environmental isolates on Earth. However, several pathogens were also isolated: Staphylococcus aureus and Escherichia coli.
Strayer, Richard F.↗
Microbial Characterization of Solid-Wastes Treated with Heat Melt Compaction Technology
The research purpose of the project was to determine the fate of microorganisms in space-generated solid wastes after processing by a Heat Melt Compactor (HMC), which is a candidate solid waste treatment technology. Five HMC product disks were generated at Ames Research Center (ARC), Waste Management Systems element. The feed for two was simulated space-generated trash and feed for three was Volume F compartment wet waste returned on STS 130. Conventional microbiological methods were used to detect and enumerate microorganisms in HMC disks and in surface swab samples of HMC hardware before and after operation. Also, biological indicator test strips were added to the STS trash prior to compaction to test if HMC processing conditions, 150 C for approx 3 hr and dehydration, were sufficient to eliminate the test bacteria on the strips. During sample acquisition at KSC, the HMC disk surfaces were sanitized with 70% alcohol to prevent contamination of disk interiors. Results from microbiological assays indicated that numbers of microbes were greatly reduced but not eliminated by the 70% alcohol. Ten 1.25 cm diameter cores were aseptically cut from each disk to sample the disk interior. The core material was run through the microbial characterization analyses after dispersal in sterile diluent. Low counts of viable bacteria (5 to 50 per core) were found but total direct counts were 6 to 8 orders of magnitude greater. These results indicate that the HMC operating conditions might not be sufficient for complete waste sterilization, but the vast majority of microbes present in the wastes were dead or non-cultivable after HMC treatment. The results obtained from analyses of the commercial spore test strips that had been added fo the wastes prior to HMC operation further indicated that the HMC was sterilizing the wastes. Nearly all strips were recovered from the HMC disks and all of these were negative for spore growth when run through the manufacturer's protocol. The 10(exp 6) or so spores impregnated into the strips were no longer viable. Control test strips, i.e., not exposed to the HMC conditions, were all strongly positive. All isolates from the cultivable counts were identified, leading to one concern: several were identified as Staphylococcus aureus, a human pathogen. The project reported here provides microbial characterization support to the Waste Management Systems element of the Life Support and Habitation Systems program.
Strayer, Richard F.↗
Microbial Characterization Space Solid Wastes Treated with a Heat Melt Compactor
The on going purpose of the project efforts was to characterize and determine the fate of microorganisms in space-generated solid wastes before and after processing by candidate solid waste processing. For FY 11, the candidate technology that was assessed was the Heat Melt Compactor (HMC). The scope included five HMC. product disks produced at ARC from either simulated space-generated trash or from actual space trash, Volume F compartment wet waste, returned on STS 130. This project used conventional microbiological methods to detect and enumerate microorganisms in heat melt compaction (HMC) product disks as well as surface swab samples of the HMC hardware before and after operation. In addition, biological indicators were added to the STS trash prior to compaction in order to determine if these spore-forming bacteria could survive the HMC processing conditions, i.e., high temperature (160 C) over a long duration (3 hrs). To ensure that surface dwelling microbes did not contaminate HMC product disk interiors, the disk surfaces were sanitized with 70% alcohol. Microbiological assays were run before and after sanitization and found that sanitization greatly reduced the number of identified isolates but did not totally eliminate them. To characterize the interior of the disks, ten 1.25 cm diameter core samples were aseptically obtained for each disk. These were run through the microbial characterization analyses. Low counts of bacteria, on the order of 5 to 50 per core, were found, indicating that the HMC operating conditions might not be sufficient for waste sterilization. However, the direct counts were 6 to 8 orders of magnitude greater, indicating that the vast majority of microbes present in the wastes were dead or non-cultivable. An additional indication that the HMC was sterilizing the wastes was the results from the added commercial spore test strips to the wastes prior to HMC operation. Nearly all could be recovered from the HMC disks post-operation and all were showed negative growth when run through the manufacturer's protocol, meaning that the 106 or so spores impregnated into the strips were dead. Control test strips, i.e., not exposed to the HMC conditions were all strongly positive. One area of concern is that the identities of isolates from the cultivable counts included several human pathogens, namely Staphylococcus aureus. The project reported here provides microbial characterization support to the Waste Management Systems element of the Life Support and Habitation Systems program.
Strayer, Richard F.↗
Efficacy of Antimicrobials on Bacteria Cultured in a Spaceflight Analogue
As humans travel in space, they will interact with microbial flora from themselves, other crewmembers, their food, and the environment. While evaluations of microbial ecology aboard the Mir and ISS suggest a predominance of common environmental flora, the presence of (and potential for) infectious agents has been well documented. Likewise, pathogens have been detected during preflight monitoring of spaceflight food, resulting in the disqualification of that production lot from flight. These environmental and food organisms range from the obligate pathogen, Salmonella enterica serovar Typhimurium (S. Typhimurium), which has been responsible for disqualification and removal of food destined for ISS and has previously been reported from Shuttle crew refuse, to the opportunistic pathogen Staphylococcus aureus, isolated numerous times from ISS habitable compartments and the crew. Infectious disease events have affected spaceflight missions, including an upper respiratory infection that delayed the launch of STS-36 and an incapacitating Pseudomonas aeruginosa urinary tract infection of a crewmember during Apollo 13. These observations indicate that the crew has the potential to be exposed to obligate and opportunistic pathogens. This risk of exposure is expected to increase with longer mission durations and increased use of regenerative life support systems. As antibiotics are the primary countermeasure after infection, determining if their efficacy during spaceflight missions is comparable to terrestrial application is of critical importance. The NASA Rotating Wall Vessel (RWV) culture system has been successfully used as a spaceflight culture analogue to identify potential alterations in several key microbial characteristics, such as virulence and gene regulation, in response to spaceflight culture. We hypothesized that bacteria cultured in the low fluid shear RWV environment would demonstrate changes in efficacy of antibiotics compared to higher fluid shear controls. This study investigated the response of three medically significant microorganisms grown in the RWV to antibiotics that could be used on spaceflight missions. Our findings suggest potential alterations in antibiotic efficacy during spaceflight and indicate that future studies on the antibiotic response require additional basic research using the RWV and/or true spaceflight. However, while this analogue has reinforced these potential alterations, the results suggest the best approach for applied forward work is evaluating an in vivo system during spaceflight, including human and rodent studies. The complex nature of the analysis for many antibiotics and organism suggests the best approach to determine in vivo responses during pharmaceutical treatment is evaluating an in vivo system during spaceflight.
Nickerson, CA↗